Recent data have demonstrated that mutations in the receptor for neurokinin B (NKB), the NK-3 receptor (NK3R), produce hypogonadotropic hypogonadism in humans. These data, together with reports that NKB expression increases after ovariectomy and in postmenopausal women, have led to the hypothesis that this tachykinin is an important stimulator of GnRH secretion. However, the NK3R agonist, senktide, inhibited LH secretion in rats and mice. In this study, we report that senktide stimulates LH secretion in ewes. A dramatic increase in LH concentrations to levels close to those observed during the preovulatory LH surge was observed after injection of 1 nmol senktide into the third ventricle during the follicular, but not in the luteal, phase. Similar increases in LH secretion occurred after insertion of microimplants containing this agonist into the retrochiasmatic area (RCh) in anestrous or follicular phase ewes. A low-dose microinjection (3 pmol) of senktide into the RCh produced a smaller but significant increase in LH concentrations in anestrous ewes. Moreover, NK3R immunoreactivity was clearly evident in the RCh, although it was not found in A15 dopaminergic cell bodies in this region. These data provide evidence that NKB stimulates LH (and presumably GnRH) secretion in ewes and point to the RCh as one important site of action. Based on these data, and the effects of NK3R mutations in humans, we hypothesize that NKB plays an important stimulatory role in the control of GnRH and LH secretion in nonrodent species.

The objectives of this study were to compare the timing and mechanisms controlling the onset of anestrus in young and mature mares treated either continuously with melatonin and in those that remained untreated. Changes in body weight, subcutaneous body fat measured to provide an estimate of total body fat, and circulating concentrations of leptin were compared throughout the 1-yr experimental period. The results demonstrate that in young mares the timing of anestrus occurs significantly earlier in the year than in mature mares and that mature mares are more likely to exhibit continuous reproductive activity during the nonbreeding season. The propensity of mature mares to exhibit this phenomenon is not modified by continuous treatment with melatonin but is associated with higher mean circulating concentrations of leptin, body weight, and estimated percent of body fat. In both young and mature mares, body weight, percent of body fat, and circulating concentrations of leptin are higher during summer than winter months. We conclude that, in the mare, the reproductive response to a decrease in photoperiod or a presumptive inhibitory melatonin signal is modified by energy availability, which may be signaled to the hypothalamus-pituitary axis via a change in the circulating concentration of leptin. An additional observation confirmed that the prolactin axis is responsive to continuous treatment with melatonin but that a suppression of prolactin secretion is limited to the spring months.

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Simple SummaryLive yeast products are used in ruminant nutrition to improve feed efficiency and performance. The effects of yeast on animal performance are mediated via alteration in the rumen microbial population and fermentation; however, the types of metabolites produced from feeding yeast additives have not been described. This study integrated 16S rRNA sequencing and LC–MS-based metabolomics to evaluate rumen bacterial diversity and metabolome of beef steers fed no or 15 g/d of live yeast product. Our findings confirm that live yeast supports the growth of fiber digesters, optimizes the utilization of oxygen and lactic acid, and inhibits the growth of pathogenic Salmonella in the rumen. In addition, some bacteria with unknown functions in relation to the effects of live yeast showed positive correlations with metabolites involved in the metabolism of amino acid and energy substrates. This study enhances our understanding of the effects of live yeast in the rumen.AbstractWe evaluated the effects of live yeast on ruminal bacterial diversity and metabolome of beef steer. Eight rumen-cannulated Holstein steers were assigned randomly to one of two treatment sequences in a study with two 25-d experimental periods and a crossover design. The steers were housed in individual pens. The dietary treatments were control (CON) or yeast (YEA; CON plus 15 g/d of live yeast product). Bacterial diversity was examined by sequencing the V3-V4 region of 16S rRNA gene. The metabolome analysis was performed using a liquid chromatograph and a mass spectrometry system (LC–MS). Live yeast supplementation increased the relative abundance of eight cellulolytic bacterial genera as well as Anaerovorax and Lachnospiraceae. Proteiniclasticum, Salmonella, and Lactococcus were not detected in the YEA treatment. Live yeast supplementation increased the concentrations of 4-cyclohexanedione and glucopyranoside and decreased the concentrations of threonic acid, xanthosine, deoxycholic acid, lauroylcarnitine, methoxybenzoic acid, and pentadecylbenzoic acid. The bacteroidales BS11, Christensenellaceae R-7, and Candidatus saccharimonas showed positive correlations with the metabolites involved in amino acid biosynthesis and the metabolism of energy substrates; the functions of these bacteria are not fully understood in relation to the mode of action of yeast. This study confirms the usefulness of LC–MS-based metabolomics in deciphering the mode of action of live yeast in the rumen.

Several studies have evaluated the effects of live yeast supplementation on rumen microbial population; however, its effect on differential microbial genes and their functional potential has not been described. Thus, this study applied shotgun metagenomic sequencing to evaluate the effects of live yeast supplementation on genetic and functional potential of the rumen microbiota in beef cattle. Eight rumen-cannulated Holstein steers were randomly assigned to two treatments in a cross-over design with two 25-day experimental periods and a 10-day wash-out between the two periods. The steers were housed in individual pens and fed 50% concentrate-mix and 50% red clover/orchard hay ad libitum . Treatments were (1) control (CON; basal diet without additive) and (2) yeast (YEA; basal diet plus 15 g/d of live yeast product). Rumen fluid samples were collected at 3, 6, and 9 h after feeding on the last d of each period. Sequencing was done on an Illumina HiSeq 2500 platform. Dietary yeast supplementation increased the relative abundance of carbohydrate-fermenting bacteria (such as Ruminococcus albus , R . champanellensis , R. bromii , and R. obeum ) and lactate-utilizing bacteria (such as Megasphaera elsdenii , Desulfovibrio desulfuricans , and D. vulgaris ). A total of 154 differentially abundant genes (DEGs) were obtained (false discovery rate < 0.01). Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation analysis of the DEGs revealed that 10 pathways, including amino sugar and nucleotide sugar metabolism, oxidative phosphorylation, lipopolysaccharide biosynthesis, pantothenate and coenzyme A biosynthesis, glutathione metabolism, beta-alanine metabolism, polyketide sugar unit biosynthesis, protein export, ribosome, and bacterial secretory system, were enriched in steers fed YEA. Annotation analysis of the DEGs in the carbohydrate-active enzymes (CAZy) database revealed that the abundance of genes coding for enzymes belonging to glycoside hydrolases, glycosyltransferases, and carbohydrate binding modules were enriched in steers fed YEA. These results confirm the effectiveness of a live S. cerevisiae product for improving rumen function in beef steers by increasing the abundance of cellulolytic bacteria, lactic acid-utilizing bacteria, and carbohydrate-active enzymes in the rumen. Electronic supplementary material The online version of this article (10.1186/s40104-019-0378-x) contains supplementary material, which is available to authorized users.

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It is now clear that seasonal breeding in ewes is due to an increase in response to estradiol (E(2)) negative feedback in the nonbreeding season (anestrus) that is mediated by the A15 group of dopaminergic (DA) neurons. Because A15 cells do not contain estrogen receptors, we have postulated the presence of estrogen-responsive afferents and recently reported evidence that input from neurons containing gamma-aminobutyric acid (GABA) contribute to the control of A15 activity by E(2). However, GABAergic afferents account for only a fraction of A15 synaptic input and do not appear to vary with season. We therefore investigated the possible role of stimulatory glutamatergic input to A15 neurons. In experiments 1 and 2, local administration into the A15 of either a N-methyl-D-aspartate (NMDA) receptor or a kainate/alpha-amino-3-hydroxyl-5-methyl-4-isoxazole-propionate (AMPA) receptor antagonist stimulated episodic LH secretion in a dose-dependent manner in ovary-intact anestrous ewes. In experiment 3, we examined the number of glutamatergic close contacts onto A15 neurons using dual immunocytochemistry in tissue from E(2)-treated ovariectomized anestrous and breeding season ewes. All A15 DA neurons were contacted by glutamatergic vesicles, and the number of close contacts was significantly higher in anestrus than the breeding season. Finally, using a triple-label immunocytochemistry procedure, we did not observe any colocalization of markers for GABA and glutamate in vesicles contacting A15 neurons. These results support the hypothesis that glutamatergic afferents actively stimulate A15 DA neurons in ovary-intact anestrous ewes and raise the possibility that alterations in this input may contribute to increased A15 neural activity during anestrus.

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